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At least 163 records · Page 9

Root angle is controlled by EGT1 in cereal crops employing an antigravitropic mechanism

Root angle in crops represents a key trait for efficient capture of soil resources. Root angle is determined by competing gravitropic versus antigravitropic offset (AGO) mechanisms. Here we report a root angle regulatory gene termed ENHANCED GRAVITROPISM1 ( EGT1 ) that encodes a putative AGO component, whose loss-of-function enhances root gravitropism. Mutations in barley and wheat EGT1 genes confer a striking root phenotype, where every root class adopts a steeper growth angle. EGT1 encodes an F-box and Tubby domain-containing protein that is highly conserved across plant species. Haplotype analysis found that natural allelic variation at the barley EGT1 locus impacts root angle. Gravitropic assays indicated that Hvegt1 roots bend more rapidly than wild-type. Transcript profiling revealed Hvegt1 roots deregulate reactive oxygen species (ROS) homeostasis and cell wall-loosening enzymes and cofactors. ROS imaging shows that Hvegt1 root basal meristem and elongation zone tissues have reduced levels. Atomic force microscopy measurements detected elongating Hvegt1 root cortical cell walls are significantly less stiff than wild-type. In situ analysis identified HvEGT1 is expressed in elongating cortical and stele tissues, which are distinct from known root gravitropic perception and response tissues in the columella and epidermis, respectively. We propose that EGT1 controls root angle by regulating cell wall stiffness in elongating root cortical tissue, counteracting the gravitropic machinery’s known ability to bend the root via its outermost tissues. We conclude that root angle is controlled by EGT1 in cereal crops employing an antigravitropic mechanism.

59 BASIC BIOLOGICAL SCIENCES↗

Expression quantitative trait loci mapping identified PtrXB38 as a key hub gene in adventitious root development in Populus

Summary Plant establishment requires the formation and development of an extensive root system with architecture modulated by complex genetic networks. Here, we report the identification of the PtrXB38 gene as an expression quantitative trait loci (eQTL) hotspot, mapped using 390 leaf and 444 xylem Populus trichocarpa transcriptomes. Among predicted targets of this trans ‐eQTL were genes involved in plant hormone responses and root development. Overexpression of PtrXB38 in Populus led to significant increases in callusing and formation of both stem‐born roots and base‐born adventitious roots. Omics studies revealed that genes and proteins controlling auxin transport and signaling were involved in PtrXB38‐mediated adventitious root formation. Protein–protein interaction assays indicated that PtrXB38 interacts with components of endosomal sorting complexes required for transport machinery, implying that PtrXB38‐regulated root development may be mediated by regulating endocytosis pathway. Taken together, this work identified a crucial root development regulator and sheds light on the discovery of other plant developmental regulators through combining eQTL mapping and omics approaches.

54 ENVIRONMENTAL SCIENCES↗

The State of the Art in Root System Architecture Image Analysis Using Artificial Intelligence: A Review

Roots are essential for acquiring water and nutrients to sustain and support plant growth and anchorage. However, they have been studied less than the aboveground traits in phenotyping and plant breeding until recent decades. In modern times, root properties such as morphology and root system architecture (RSA) have been recognized as increasingly important traits for creating more and higher quality food in the “Second Green Revolution”. To address the paucity in RSA and other root research, new technologies are being investigated to fill the increasing demand to improve plants via root traits and overcome currently stagnated genetic progress in stable yields. Artificial intelligence (AI) is now a cutting-edge technology proving to be highly successful in many applications, such as crop science and genetic research to improve crop traits. A burgeoning field in crop science is the application of AI to high-resolution imagery in analyses that aim to answer questions related to crops and to better and more speedily breed desired plant traits such as RSA into new cultivars. This review is a synopsis concerning the origins, applications, challenges, and future directions of RSA research regarding image analyses using AI.

59 BASIC BIOLOGICAL SCIENCES↗

Electrical capacitance as a predictor of root dry weight in shrub willow ( Salix ; Salicaceae) parents and progeny

Premise of the study: Root biomass is an important trait often disregarded in woody perennial selection due to the challenge and expense of accurately and efficiently measuring large populations. In this study, we aim to develop a simple method that can predict root dry weight within a diverse shrub willow ( Salix ) breeding population representing species hybrids and their parents using root electrical capacitance (REC). Methods: The REC method was tested on plants started from cuttings and grown in pots with potting mix in the greenhouse for 11 wk to assess the relationship of REC with 24 biomass traits and its usefulness in allometric models for root and stem dry biomass. Results: Strong linear and positive correlations were found between REC and root dry biomass ( r = 0.88). The total proportion of variance of root and stem dry biomass explained by predictors in multiple regression was 85% and 69%, respectively. The relative importance of predictor variables in allometric models was dominated by the contribution of REC. Discussion: This work provides an efficient and nondestructive technique to indirectly quantify root biomass of genetically diverse shrub willow progeny, which has great promise for selection of genotypes with varying root biomass and for the accurate estimation of belowground carbon sequestration.

59 BASIC BIOLOGICAL SCIENCES↗

Metabotyping as a Stopover in Genome-to-Phenome Mapping

Predicting phenotypic expression from genomic and environmental information is arguably the greatest challenge in today’s biology. Being able to survey genomic content, e.g., as singlenucleotide polymorphism data, within a diverse population and predict the phenotypes of external traits, represents the holy grail across genome-informed disciplines, from personal medicine and nutrition to plant breeding. In the present study, we propose a two-step procedure in bridging the genome to phenome gap where external phenotypes are viewed as emergent properties of internal phenotypes, such as molecular profiles, in interaction with the enviorment. Using biomass accumulation and shoot-root allometry as external traits in diverse genotypes of the model grass Brachypodium distachyon , we established correlative models between genotypes and metabolite profiles (metabotypes) as internal phenotypes, and between metabotypes and external phenotypes under two contrasting watering regimes. Our results demonstrate the potential for employing metabotypes as an integrator in predicting external phenotypes from genomic information.

59 BASIC BIOLOGICAL SCIENCES↗

GWAS identifies candidate genes controlling adventitious rooting in Populus trichocarpa

Adventitious rooting (AR) is critical to the propagation, breeding, and genetic engineering of trees. The capacity for plants to undergo this process is highly heritable and of a polygenic nature; however, the basis of its genetic variation is largely uncharacterized. To identify genetic regulators of AR, we performed a genome-wide association study (GWAS) using 1148 genotypes of Populus trichocarpa. GWASs are often limited by the abilities of researchers to collect precise phenotype data on a high-throughput scale; to help overcome this limitation, we developed a computer vision system to measure an array of traits related to adventitious root development in poplar, including temporal measures of lateral and basal root length and area. GWAS was performed using multiple methods and significance thresholds to handle non-normal phenotype statistics and to gain statistical power. These analyses yielded a total of 277 unique associations, suggesting that genes that control rooting include regulators of hormone signaling, cell division and structure, reactive oxygen species signaling, and other processes with known roles in root development. Numerous genes with uncharacterized functions and/or cryptic roles were also identified. These candidates provide targets for functional analysis, including physiological and epistatic analyses, to better characterize the complex polygenic regulation of AR.

59 BASIC BIOLOGICAL SCIENCES↗

Data from TropiRoot 1.0 database: tropical root characteristics across environments

TropiRoot 1.0 is a new tropical root database with root characteristics across environment gradients. It has data extracted from 104 new sources, resulting in more than 8000 rows of data (either species or community data). Most of the data in TropiRoot 1.0 includes root characteristics such as root biomass, morphology, root dynamics, mass fraction, architecture, anatomy, physiology and root chemistry. This initiative represents an approximately 30% increase in the currently available data for tropical roots in the Fine Root Ecology Database (FRED). TropiRoot 1.0, contains root characteristics from 25 different countries where seven are located in Asia, six in South America, five in Central America and the Caribbean, four in Africa, two in North America, and 1 in Oceania. Due to the volume of data, when ancillary data was available, including soil data, these data was either extracted and included in the database or their availability was recorded in an additional column. Multiple contributors checked the entries for outliers during the collation process to ensure data quality. For text-based observations, we examined all cells to ensure that their content relates to their specific categories. For numerical observations, we ordered each numerical value from least to greatest and plotted the values, checking apparent outliers against the data in their respective sources and correcting or removing incorrect or impossible values. Some data (soil and aboveground) have different columns for the same variable presented in different units, including originally published units, but root characteristics data had units converted to match the ones reported in FRED. By filling a gap from global databases, TropiRoot 1.0 expands our knowledge of otherwise so far underrepresented regions, and our ability to assess global trends. This advancement can be used to improve tropical forest representation in vegetation models.

54 ENVIRONMENTAL SCIENCES↗

Root and Rhizosphere Bacterial Phosphatase Activity Varies with Tree Species and Soil Phosphorus Availability in Puerto Rico Tropical Forest

Climatic conditions in tropical forests combined with the immobility of phosphorus due to sorption on mineral surfaces or result in soils typically lacking in the form of phosphorus (orthophosphate) most easily metabolized by plants and microbes. In these soils, mineralization of organic phosphorus can be the major source for labile inorganic P available for uptake. Both plants and microbes encode for phosphatase enzymes capable of mineralizing a range of organic phosphorus compounds. However, the activity of these enzymes depends on several edaphic factors including P availability and tree or microbial species. Thus, phosphatase activity in both roots and the root microbial community constitute an important role in P mineralization and P nutrient dynamics that are not well studied in tropical forests. We measured phosphatase activity in roots and bacterial isolates from the microbial community of six tree species from three forest sites differing in phosphorus availability in the Luquillo Mountains of Puerto Rico. Root and microbial phosphatase activity were both influenced by tree identity and soil phosphorus availability. However, tree identity had a larger effect on phosphatase activity (effect size = 0.12) than soil phosphorus availability (effect size = 0.07). In addition, lower amounts of P availability corresponded with higher levels of enzyme activity. In contrast, ANOSIM analysis of the weighted UniFrac distance matrix indicates that microbial community composition was more strongly controlled by soil P availability (P value < 0.05). These results indicate that root and rhizosphere microbial phosphatase activity are similarly expressed despite the slightly stronger influence of tree identity on root function and the stronger influence of P availability on microbial community composition. The low levels of orthophosphate in tropical forests, rather than prohibiting growth, have encouraged a variety of functions to adapt to minimal levels of an essential nutrient. Phosphatase activity is one such mechanism that varies in both roots and microbial community members. A thorough understanding of phosphatase activity provides insight into P mineralization in tropical forests, providing not only perspective on ecosystem function of tropical trees and microbial communities, but also in advancing efforts to improve representations of tropical forests in future climates.

54 ENVIRONMENTAL SCIENCES↗

Filling gaps in our understanding of belowground plant traits across the world: an introduction to a Virtual Issue

The belowground world is one of the final frontiers in terrestrial ecology. The tangling of plant roots with the surrounding soil below is a lifeline for the humble forbs and towering trees above, and roots play a key role in shaping ecosystem carbon, water and nutrient cycling (Bardgett et al., 2014). Ecologists have long sought to better understand the ecosystem-scale consequences of differing plant strategies, above- and belowground, by relating plant characteristics, or traits, to plant function (Grime, 1977; Pregitzer, 2002). While developing trait–function linkages is arguably more difficult for plant traits that are hidden belowground, root and rhizosphere ecologists continue to fan out across grasslands and forests with their shovels, isotopes, and specialized cameras, seeking a better understanding of the secret lives of roots. Over the years, New Phytologist has served as a virtual town square for scientists to discuss their hard-won observations on the interplay among belowground plant traits, microbial activity, and edaphic and environmental conditions from biomes around the world (Norby & Jackson, 2000; Pregitzer, 2002; Matamala & Stover, 2013; Norby & Iversen, 2017). In this Editorial we highlight the newest papers that update and add to our understanding of the role of root and rhizosphere traits in broader ecosystem processes. We focused on papers published in New Phytologist between 1 January 2019 and 31 December 2020 and extended this window to papers still in ‘early view’ up to the time of writing. Because of the overwhelming number of papers, we did not include those with a decidedly genomic focus or that served primarily as data syntheses, reviews, insights or commentaries.

59 BASIC BIOLOGICAL SCIENCES↗

Relatively rare root endophytic bacteria drive plant resource allocation patterns and tissue nutrient concentration in unpredictable ways

Plant endophytic bacterial strains can influence plant traits such as leaf area and root length. Yet, the influence of more complex bacterial communities in regulating overall plant phenotype is less explored. Here, in two complementary experiments, we tested whether we can predict plant phenotype response to changes in microbial community composition. In the first study, we inoculated a single genotype of Populus deltoides with individual root endophytic bacteria and measured plant phenotype. Next, data from this single inoculation were used to predict phenotypic traits after mixed three-strain community inoculations, which we tested in the second experiment. By itself, each bacterial endophyte significantly but weakly altered plant phenotype relative to noninoculated plants. In a mixture, bacterial strain Burkholderia BT03, constituted at least 98% of community relative abundance. Yet, plant resource allocation and tissue nutrient concentrations were disproportionately influenced by Pseudomonas sp. GM17, GM30, and GM41. We found a 10% increase in leaf mass fraction and an 11% decrease in root mass fraction when replacing Pseudomonas GM17 with GM41 in communities containing both Pseudomonas GM30 and Burkholderia BT03. Our results indicate that interactions among endophytic bacteria may drive plant phenotype over the contribution of each strain individually. Additionally, we have shown that low-abundance strains contribute to plant phenotype challenging the assumption that the dominant strains will drive plant function.

59 BASIC BIOLOGICAL SCIENCES↗

Global relationships in tree functional traits

Due to massive energetic investments in woody support structures, trees are subject to unique physiological, mechanical, and ecological pressures not experienced by herbaceous plants. Despite a wealth of studies exploring trait relationships across the entire plant kingdom, the dominant traits underpinning these unique aspects of tree form and function remain unclear. Here, by considering 18 functional traits, encompassing leaf, seed, bark, wood, crown, and root characteristics, we quantify the multidimensional relationships in tree trait expression. We find that nearly half of trait variation is captured by two axes: one reflecting leaf economics, the other reflecting tree size and competition for light. Yet these orthogonal axes reveal strong environmental convergence, exhibiting correlated responses to temperature, moisture, and elevation. By subsequently exploring multidimensional trait relationships, we show that the full dimensionality of trait space is captured by eight distinct clusters, each reflecting a unique aspect of tree form and function. Collectively, this work identifies a core set of traits needed to quantify global patterns in functional biodiversity, and it contributes to our fundamental understanding of the functioning of forests worldwide.

54 ENVIRONMENTAL SCIENCES↗

Uncovering natural variation in root system architecture and growth dynamics using a robotics-assisted phenomics platform

The plant kingdom contains a stunning array of complex morphologies easily observed above-ground, but more challenging to visualize below-ground. Understanding the magnitude of diversity in root distribution within the soil, termed root system architecture (RSA), is fundamental in determining how this trait contributes to species adaptation in local environments. Roots are the interface between the soil environment and the shoot system and therefore play a key role in anchorage, resource uptake, and stress resilience. Previously, we presented the GLO-Roots (Growth and Luminescence Observatory for Roots) system to study the RSA of soil-grown Arabidopsis thaliana plants from germination to maturity (Rellán-Álvarez et al., 2015). In this study, we present the automation of GLO-Roots using robotics and the development of image analysis pipelines in order to examine the temporal dynamic regulation of RSA and the broader natural variation of RSA in Arabidopsis , over time. These datasets describe the developmental dynamics of two independent panels of accessions and reveal highly complex and polygenic RSA traits that show significant correlation with climate variables of the accessions’ respective origins.

59 BASIC BIOLOGICAL SCIENCES↗

Perennial grass root system specializes for multiple resource acquisitions with differential elongation and branching patterns

Roots optimize the acquisition of limited soil resources, but relationships between root forms and functions have often been assumed rather than demonstrated. Furthermore, how root systems co-specialize for multiple resource acquisitions is unclear. Theory suggests that trade-offs exist for the acquisition of different resource types, such as water and certain nutrients. Measurements used to describe the acquisition of different resources should then account for differential root responses within a single system. To demonstrate this, we grew Panicum virgatum in split-root systems that vertically partitioned high water availability from nutrient availability so that root systems must absorb the resources separately to fully meet plant demands. We evaluated root elongation, surface area, and branching, and we characterized traits using an order-based classification scheme. Plants allocated approximately 3/4th of primary root length towards water acquisition, whereas lateral branches were progressively allocated towards nutrients. However, root elongation rates, specific root length, and mass fraction were similar. Our results support the existence of differential root functioning within perennial grasses. Similar responses have been recorded in many plant functional types suggesting a fundamental relationship. Root responses to resource availability can be incorporated into root growth models via maximum root length and branching interval parameters.

59 BASIC BIOLOGICAL SCIENCES↗

Hydraulic anatomy affects genotypic variation in plant water use and shows differential organ specific plasticity to drought in Sorghum bicolor

Genotypic variability and plasticity in hydraulic anatomy are not well-understood in herbaceous monocots. In this work, we used Sorghum bicolor , a monocotyledonous, tropical grass model, to understand whether differential plant water use is associated with xylem anatomy and if whole-plant xylem anatomy responds to water stress, justifying differential genotypic sensitivity to drought. In a greenhouse environment, we studied four sorghum genotypes that are known to genetically differ in growth and exhibit differential sensitivity to drought. Under well-watered scenario, transpiration variability and plant growth traits correlated with xylem anatomical traits at both the leaf and stem level, including xylem area and predicted xylem-specific hydraulic conductivity. High water use genotypes had inherently higher hydraulic capacity, but under drought, their transpiration declined at higher fractions of transpirable soil water ( FTSW ) and they showed greater plasticity in hydraulic anatomy. However, lower FTSW thresholds and modest anatomical changes were identified in the low water use genotypes with inherently lower hydraulic conductivity. Drought, induced modular phenotypic plasticity in hydraulic anatomy, whereby plasticity in leaf xylem traits was remarkably higher than stem xylem, while root xylem showed a reverse nature of vascular modification. Xylem traits were in agreement with phloem anatomy, irrespective of water regime. In conclusion, our study indicates that hydraulic anatomy can be critical for herbaceous monocots in determining limits to plant water use and genotypic response to drought with implications on whole-plant functions and habitat ecology.

54 ENVIRONMENTAL SCIENCES↗

Testing mechanisms of how mycorrhizal associations affect forest soil carbon and nitrogen cycling (Final Technical Report)

Trees are in a symbiotic partnership with mycorrhizal fungi in which they provide the fungi with carbon from photosynthesis and the fungi provide the trees with nutrients and water. In temperate forests, the vast majority of trees form symbioses with one of two types of mycorrhizal fungi—arbuscular mycorrhizal (AM) fungi or ectomycorrhizal (EcM) fungi. These fungi differ in their morphology, hyphal length, and nutrient acquisition strategies. Many studies have found systematic differences in soil organic matter and nitrogen availability between forest stands dominated by AM-associating trees versus EcM-associating trees. For instance, there is a larger proportion of organic matter that is mineral-associated, more available nitrogen, and lower soil carbon to nitrogen ratios in AM forest stands relative to EcM forests stands. However, the mechanisms driving these patterns are not known, which complicates our ability to model soil organic matter dynamics in forested ecosystems. The main objective of this research was to understand the degree to which the observed differences in soil C and N dynamics between AM and EcM dominated forests are driven by tree traits like litter decomposability and root exudation versus mycorrhizal fungal nutrient acquisition strategies. We investigated these mechanisms using observations and targeted experiments and incorporated this knowledge into a process-based soil organic matter model. The observational studies compared the importance of leaf litter decomposability versus fungal identity on soil organic matter processes. We found that often fungal identity and traits were more important drivers of soil organic matter patterns than leaf litter decomposability. We ran two novel experiments: 1) a growth chamber experiment across four EcM and four AM tree species using a 13 C-labeled atmosphere to trace seedling-derived C into hyphae, the rhizosphere, and soil; and 2) an in situ decomposition experiment of six different 13 C and 15 N labeled litters that ranged in decomposability incubated across a gradient of EcM dominance at three sites that capture important variation in climate, soils, and forest species composition. In the first experiment, we found no significant differences of seedling mycorrhizal association on soil carbon sequestration over a growing season, but we did find that mycorrhizal association affected rhizodeposition with EcM-associating seedlings depositing more carbon in response to increased nitrogen availability. The decomposition experiment is still ongoing, but thus far, we have found slower litter decomposition in only one of three EcM-dominated forests which suggests that differences between AM- and EcM-dominated forests depend on the environmental context and identity of the EcM fungi. Lastly, we explicitly incorporated mycorrhizal processes into the Carbon, Organisms, Rhizosphere, and Protection in the Soil Environment (CORPSE) model creating Myco-CORPSE. By including the different nutrient acquisition strategies of AM and EcM fungi, we explored the conditions under which EcM fungi can slow decomposition rates and lead to greater soil organic carbon accumulation compared to AM fungi. We found that the effect of EcM fungi was highly context dependent and that EcM fungi decreased decomposition in colder forests with recalcitrant litter inputs and when they produced oxidases and necromass-degrading enzymes. Our research highlights the importance of fungal nutrient acquisition in driving soil organic matter patterns and the need to move beyond the AM-EcM dichotomy to consider the identity and traits of the specific fungi participating in the symbiosis. Overall, this research has resulted in six, peer-reviewed published papers in journals such as Global Change Biology, Ecology (2), Soil Biology and Biochemistry, and Ecosystems (2). There are at least two more papers in progress on this research including one that was recently submitted to Global Change Biology.

54 ENVIRONMENTAL SCIENCES↗

It's only natural: Plant respiration in unmanaged systems

Abstract Respiration plays a key role in the terrestrial carbon cycle and is a fundamental metabolic process in all plant tissues and cells. We review respiration from the perspective of plants that grow in their natural habitat and how it is influenced by wide-ranging elements at different scales, from metabolic substrate availability to shifts in climate. Decades of field-based measurements have honed our understanding of the biological and environmental controls on leaf, root, stem, and whole-organism respiration. Despite this effort, there remain gaps in our knowledge within and across species and ecosystems, especially in more challenging-to-measure tissues like roots. Recent databases of respiration rates and associated leaf traits from species representing diverse biomes, plant functional types, and regional climates have allowed for a wider-lens view at modeling this important CO2 flux. We also re-analyze published data sets to show that maximum leaf respiration rates (R max) in species from around the globe are related both to leaf economic traits and environmental variables (precipitation and air temperature), but that root respiration does not follow the same latitudinal trends previously published for leaf data. We encourage the ecophysiological community to continue to expand their study of plant respiration in tissues that are difficult to measure and at the whole plant and ecosystem levels to address outstanding questions in the field.

59 BASIC BIOLOGICAL SCIENCES↗

Balancing growth amidst salt stress – lifestyle perspectives from the extremophyte model Schrenkiella parvula

Schrenkiella parvula, a leading extremophyte model in Brassicaceae, can grow and complete its lifecycle under multiple environmental stresses, including high salinity. Yet, the key physiological and structural traits underlying its stress-adapted lifestyle are unknown along with trade-offs when surviving salt stress at the expense of growth and reproduction. We aimed to identify the influential adaptive trait responses that lead to stress-resilient and uncompromised growth across developmental stages when treated with salt at levels known to inhibit growth in Arabidopsis and most crops. Its resilient growth was promoted by traits that synergistically allowed primary root growth in seedlings, the expansion of xylem vessels across the root-shoot continuum, and a high capacity to maintain tissue water levels by developing thicker succulent leaves while enabling photosynthesis during salt stress. A successful transition from vegetative to reproductive phase was initiated by salt-induced early flowering, resulting in viable seeds. Self-fertilization in salt-induced early flowering was dependent upon filament elongation in flowers otherwise aborted in the absence of salt during comparable plant ages. Furthermore, the maintenance of leaf water status promoting growth, and early flowering to ensure reproductive success in a changing environment, were among the most influential traits that contributed to the extremophytic lifestyle of S. parvula.

59 BASIC BIOLOGICAL SCIENCES↗